Gut Microbiota In Piglets Research Articles

Heat-Killed Lactobacillus acidophilus Promotes Growth by Modulating the Gut Microbiota Composition and Fecal Metabolites of Piglets.

Probiotics can improve animal growth performance and intestinal health. However, understanding the effects of paraprobiotics on the growth performance and gut microbiota of piglets and how the paraprobiotics exert their impact are still limited. The present study was conducted to investigate the effects of heat-killed Lactobacillus acidophilus IFFI 6005 supplementation on the growth performance, intestinal microbiota, and fecal metabolites of piglets. First, a feed-additive sample of heat-killed Lactobacillus acidophilus IFFI 6005 was prepared by culture. Second, 96 (initial BW = 14.38 ± 0.67 kg, weaning age of 40 days) healthy piglets were selected and randomized into four treatment groups. Each treatment group consisted of three replicates (n = 8). Pigs were fed a basal diet (NC), basal diet plus antibiotics (PC), basal diet plus Lactobacillus acidophilus IFFI 6005 at 600 g/t (LA, 1.0 × 1010 cfu/g), and basal diet plus heat-killed Lactobacillus acidophilus IFFI 6005 at 600 g/t (HKLA), respectively; the trial lasted for 30 days. The results showed that the ratios of feed to gain (F:G) and diarrhea rate of both the HKLA and PC groups were significantly lower compared with the NC and LA groups (p < 0.05); however, there was no significant difference between the HKLA and PC group (p > 0.05). In addition, the average daily weight gain (ADG) of the HKLA group was significantly higher (p < 0.05) than that of the other three groups in terms of growth performance. Finally, 16S rRNA sequencing and metabolome analysis based on fecal samples further elaborated that the addition of heat-killed Lactobacillus acidophilus IFFI 6005 to the feed improved the intestinal microbial diversity and abundance (p < 0.05) and reduced the abundance of pathogenic bacteria (p < 0.05), but it did not affect the abundance of Lactobacillus (p > 0.05). Through the comparison of microbial abundance and metabolite content between the two groups (NC_vs_HKLA), the largest differences were found in six microorganisms and 10 metabolites in the intestine (p < 0.05). These differential metabolites were involved in the digestion, absorption and utilization of protein and starch, as well as in oxidative stress. In summary, addition of heat-killed Lactobacillus acidophilus IFFI 6005 as a new feed additive in piglets has beneficial effects on the growth performance, intestinal bacteria and metabolites, and can be used as an alternative to antibiotics.

Precise evaluation of the nutritional value of yeast culture and its effect on pigs fed low-protein diets

The purpose of the present study was to assess the nutritional value of yeast culture (YC) and to explore the effect of YC on growth performance and health of piglets fed low-protein (LP) diets. In Exp.1, 12 growing barrows were allocated into control diet and YC diet treatments to determine the available energy of YC. Results showed that the digestible energy and metabolizable energy of YC are 12.12 and 11.66 MJ/kg dry matter (DM), respectively. In Exp.2, 12 growing barrows were surgically equipped with a T-cannula near the distal ileum and were assigned to 2 dietary treatments (nitrogen-free diet and YC diet), and the amino acid digestibility of YC was determined. In Exp.3, a total of 96 weaned piglets were randomly divided into 4 treatments, including LP basal diet (Basal), Basal+0.5% YC (0.5%YC), Basal+1.0% YC (1%YC), and Basal+1.5% YC (1.5%YC). The results were as follows: YC supplementation linearly improved the weight gain and feed intake ratio (P < 0.001), linearly increased the activity of glutathione peroxidase on d 14 (P = 0.032) and linearly decreased the concentration of malondialdehyde on d 14 (P = 0.008) and d 32 (P = 0.004) in serum, and linearly decreased the concentration of total short-chain fatty acid on d 14 in feces (P = 0.045). Compared with other treatments, 1.5%YC group showed a greater abundance of various probiotics, such as Prevotellaceae, Prevotella and Turicibacter. In Exp.4, twelve growing barrows with an ileal T-cannula were randomly assigned to Control and 1.5%YC treatments to clarify the impact of YC supplementation on nitrogen balance and nutrient digestibility. Results showed that YC had no significant effect on nitrogen efficiency and nutrient digestibility, except for trend of reducing the total tract digestibility of organic matter (P = 0.067). In conclusion, the present study assessed the digestible and metabolizable energy values (12.12 and 11.66 MJ/kg DM, respectively) and standardized ileal digestibility of amino acid (from 43.93% to 82.65%) of YC in pig feed and demonstrated that moderate supplementation of YC (1.5% of diet) can effectively improve feed conversion efficiency, enhance antioxidant capacity, and promote a balanced gut microbiota in piglets.

Ningxiang pig-derived Enterococcus hirae HNAU0516 ameliorates postweaning diarrhoea by promoting intestinal health and modulating the gut microbiota in piglets

Early weaning-induced stress precipitates diarrhoea, significantly curtailing the growth performance of piglets. A pivotal contributor to this postweaning affliction is the emergence of gut bacterial dysbiosis. Enterococcus hirae, a promising probiotic, has indicated unclear effects and mechanisms on intestinal health. In this study, we investigated the effects and underlying mechanisms of oral supplementation with Ningxiang pig-derived Enterococcus hirae HNAU0516 orally supplementation on the gut bacterial community, immune response and gut barrier function in piglets. 21 d age Duroc × (Landrace × Yorkshire) piglets with a similar BW were randomly allocated to two groups. The Enterococcus hirae HNAU0516 administration group was inoculated orally with Ningxiang pig-derived Enterococcus hirae HNAU0516 throughout the trial period. Conversely, the control group received the same volume of physiological saline. Our findings revealed that Enterococcus hirae HNAU0516 supplementation effectively reduced diarrhoea rates of piglets (P = 0.010). Notably, this probiotic promoted intestinal development and enhanced intestinal barrier function. It also showed potential anti-inflammatory properties. Furthermore, Enterococcus hirae HNAU0516 supplementation significantly remodelled the colonic microbiota and increased the production of acetate (P = 0.007). In conclusion, our study highlights that Ningxiang pig-derived Enterococcus hirae HNAU0516 improves postweaning diarrhoea by promoting intestinal development, enhancing intestinal barrier function, decreasing intestinal permeability, modulating intestinal microbiota, and increasing short-chain fatty acids production.

Protecting the piglet gut microbiota against ETEC-mediated post-weaning diarrhoea using specific binding proteins

Post-weaning diarrhoea (PWD) in piglets presents a widespread problem in industrial pig production and is often caused by enterotoxigenic E. coli (ETEC) strains. Current solutions, such as antibiotics and medicinal zinc oxide, are unsustainable and are increasingly being prohibited, resulting in a dire need for novel solutions. Thus, in this study, we propose and evaluate a protein-based feed additive, comprising two bivalent heavy chain variable domain (VHH) constructs (VHH-(GGGGS)3-VHH, BL1.2 and BL2.2) as an alternative solution to manage PWD. We demonstrate in vitro that these constructs bind to ETEC toxins and fimbriae, whilst they do no affect bacterial growth rate. Furthermore, in a pig study, we show that oral administration of these constructs after ETEC challenge reduced ETEC proliferation when compared to challenged control piglets (1-2 log10 units difference in gene copies and bacterial count/g faeces across day 2–7) and resulted in week 1 enrichment of three bacterial families (Prevotellaceae (estimate: 1.12 ± 0.25, q = 0.0054), Lactobacillaceae (estimate: 2.86 ± 0.52, q = 0.0012), and Ruminococcaceae (estimate: 0.66 ± 0.18, q = 0.049)) within the gut microbiota that appeared later in challenged control piglets, thus pointing to an earlier transition towards a more mature gut microbiota. These data suggest that such VHH constructs may find utility in industrial pig production as a feed additive for tackling ETEC and reducing the risk of PWD in piglet populations.

Limosilactobacillus reuteri alleviates weaned stress by improving immune function and gut microbiota in piglets

Weaned stress may damage the tissue development, barrier function, and balance of microbiota of gut, resulting in the diarrhea and reduced growth performance of piglets. Probiotics with excellent characteristics can enhance the resistance of piglets to weaned stress. Under a poor sanitary condition, the Limosilactobacillus reuteri (L. reuteri) SLZX19-12 was supplemented to check its effects for piglets. Results showed that supplementation of L. reuteri significantly decreased the diarrhea incident, improved the growth performance, and enhanced the barrier integrity of gut. In ileum, L. reuteri decreased inflammation by NFκB p65 pathway, increased propionate, TCDCA, and THDCA contents, and regulated microbiota. In colon, L. reuteri decreased the inflammation by NFκB p65 pathway, reduced DCA content, and regulated microbiota. Therefore, the supplementation of L. reuteri may alleviate weaned stress by improving immune function and regulating the gut microbiota in piglets.

The Effect of Sodium Alginate-Coated Nano-Zinc Oxide on the Growth Performance, Serum Indexes and Fecal Microbial Structure of Weaned Piglets.

High dose of zinc oxide (ZnO) could improve growth performance and alleviate disease status, whereas it caused serious environmental pollution and bacterial resistance. This study was to investigate whether low doses of sodium alginate-coated nano zinc oxide (saZnO), a new type of zinc resource, could serve as a potential alternative to pharmacological doses of traditional ZnO in weaned piglets. A total of 144 crossbred piglets were randomly allocated into three groups, including a basal diet without the addition of Zn (CON), a basal diet with 1600 mg Zn/kg from traditional ZnO (ZnO), and a basal diet with 500 mg Zn/kg from saZnO (saZnO). The experiment lasted for 28 days. The results showed that supplementing with ZnO and saZnO for 14 and 28 days significantly improved body weight (BW) and average daily gain (ADG) (p < 0.01) and markedly reduced the feed intake-to-gain ratio (F/G) (p < 0.05) and diarrhea rate. In addition, dietary ZnO and saZnO significantly increased the activities of the total antioxidant capacity (T-AOC) and alkaline phosphatase (ALP) (p < 0.01). Supplementing with saZnO also promoted the levels of superoxide dismutase (SOD), IgM and copper- and zinc-containing superoxide dismutase (Cu/Zn-SOD) in serum (p < 0.05), whereas a ZnO addition decreased the concentration of malondialdehyde (MDA) (p < 0.05), indicating the beneficial effect of Zn on antioxidant and immune functions. Piglets fed the ZnO diet showed higher serum Zn accumulations than those fed the CON and saZnO diets at d 28 (p < 0.01), and supplementing with ZnO and saZnO markedly contributed to Zn excretion in feces, especially in the ZnO diet (p < 0.01). Additionally, piglets fed the saZnO diet had greater valeric acid concentrations (p < 0.05) in their feces, while other short chain fatty acids (SCFAs) were not affected by different treatments (p > 0.05). Microbial alpha diversity was reduced in the saZnO group compared with the CON group (p < 0.05), while an obvious separation of microbial composition, the marker of beta diversity, was shown among the three groups (p < 0.05). At the genus level, six genera, including Clostridium_sensu_stricto_1, Terrisporobacter, f_Muribaculaceae, Subdoligranulum and Intestinibacter, were pronouncedly increased in the ZnO and saZnO groups (p < 0.05); another nine species were dramatically downregulated, such as f_Lachnospiraceae, f_Prevotellaceae, f_Butyricicoccaceae and f_Ruminococcaceae (p < 0.05). Finally, a functional analysis indicated that altered microbes significantly changed the "Metabolism" pathway (p < 0.05). These findings suggested that saZnO could act as a feasible substitute for ZnO to reduce Zn emission and enhance growth performance, antioxidant and immune functions, and to adjust the structure of gut microbiota in piglets.

Composition, function, and timing: exploring the early-life gut microbiota in piglets for probiotic interventions

BackgroundThe establishment of a robust gut microbiota in piglets during their early developmental stage holds the potential for long-term advantageous effects. However, the optimal timeframe for introducing probiotics to achieve this outcome remains uncertain.ResultsIn the context of this investigation, we conducted a longitudinal assessment of the fecal microbiota of 63 piglets at three distinct pre-weaning time points. Simultaneously, we gathered vaginal and fecal samples from 23 sows. Employing 16S rRNA gene and metagenomic sequencing methodologies, we conducted a comprehensive analysis of the fluctuation patterns in microbial composition, functional capacity, interaction networks, and colonization resistance within the gut microbiota of piglets. As the piglets progressed in age, discernible modifications in intestinal microbial diversity, composition, and function were observed. A source-tracking analysis unveiled the pivotal role of fecal and vaginal microbiota derived from sows in populating the gut microbiota of neonatal piglets. By D21, the microbial interaction network displayed a more concise and efficient configuration, accompanied by enhanced colonization resistance relative to the other two time points. Moreover, we identified three strains of Ruminococcus sp. at D10 as potential candidates for improving piglets' weight gain during the weaning phase.ConclusionsThe findings of this study propose that D10 represents the most opportune juncture for the introduction of external probiotic interventions during the early stages of piglet development. This investigation augments our comprehension of the microbiota dynamics in early-life of piglets and offers valuable insights for guiding forthcoming probiotic interventions.

Effects of Different Types of Dietary Fibers on Lipid Metabolism and Bile Acids in Weaned Piglets.

The aim of this study was to investigate the effects of dietary fiber on the serum biochemistry, bile acid profile, and gut microbiota in piglets. Twenty-four pigs (initial body weight: 10.53 ± 1.23 kg) were randomly divided into three treatments with eight replicate pens of one pig per pen for 21 d. The dietary treatments consisted of the following: (1) a fiber-free diet (NS); (2) a fiber-free diet + 3% fructooligosaccharides (SI); (3) a fiber-free diet + 3% dietary fiber mixture (fructooligosaccharides, long-chain inulin, and microcrystalline cellulose at the ratio 1:1:1; MIX). The results showed that compared with the NS group, the 3% SI diet reduced the serum total cholesterol (TC) concentration of the piglets (p < 0.05). The metabolomics results showed that the 3% SI diet increased the level of taurohyocholic acid (THCA) and α-muricholic acid, and the 3% MIX diet increased the level of THCA and cholic acid (p < 0.05). The use of 3% SI or MIX decreased the glycodeoxycholic acid (GDCA) level in the bile of the piglets (p < 0.05). The correlation analysis shows that the GDCA was positively related to the TC. The 16S rRNA gene sequencing results showed that UCG-002 and Holdemanella were enriched in the SI group, while Bacteroides was enriched in the MIX group. The microbial function prediction indicated that SI supplementation tended to elevate the relative abundance of gut bacteria capable of expressing bile acid-metabolizing enzymes. To sum up, the regulatory effect of dietary fiber on lipid metabolism is related to bile acids in piglets. Compared with MIX, SI is more likely to regulate bile acids through the gut microbiota.

Dietary emodin alleviates lipopolysaccharide-induced intestinal mucosal barrier injury by regulating gut microbiota in piglets.

This study was to determine the effects of dietary emodin (ED) on the intestinal mucosal barrier, nuclear factor kappa-B (NF-κB) pathways, and gut microbial flora in lipopolysaccharide (LPS)-induced piglets. Twenty-four weaned piglets were chosen and 4 treatments were created by randomly distributing piglets into CON, ED, LPS, and ED_LPS groups. Experiments were done in a 2×2 factorial arrangement and maintained for 21d. Dietary treatment (a basal diet or 300mg/kg ED) and immunological challenge (LPS or sterile saline) were 2 major factors. Intraperitoneal injections of LPS or sterilized saline were given to piglets on d 21. Six hours after the LPS challenge, all piglets were euthanized for sample collection and analysis. The results showed that piglets of the ED_LPS group had higher (P<0.05) villus height to crypt depth ratio (VCR), and lower (P<0.05) plasma D-lactate and diamine oxidase (DAO) than the LPS group. Furthermore, ED inhibited (P<0.05) the decrease of glutathione peroxidase (GSH-Px) and catalase (CAT) activities and increase of malonaldehyde level (P<0.05) in jejunal mucosa induced by LPS. The mRNA levels of pro-inflammatory cytokine genes (IL-6, IL-1β, and TNF-α) were significantly reduced (P<0.05), and the mRNA levels of antioxidant enzyme genes (GPX-1, SOD2 and CAT), as well as protein and mRNA levels of tight junction proteins (occludin, claudin-1, and ZO-1), were also significantly increased (P<0.05) by ED addition in LPS-induced piglets. Meanwhile, ED supplementation significantly decreased the LPS-induced protein levels of cyclooxygenase-2 and phosphorylation levels of NF-κB p65 and IκBα in jejunal mucosa. Emodin had a significant effect on the composition of gut microbial flora at various taxonomic positions as indicated by 16S RNA sequencing. The acetic acid, isobutyric acid, valeric acid, and isovaleric acid concentrations in the cecum were also increased by ED addition in pigs (P<0.05). Furthermore, the correlation analysis revealed that some intestinal microbiota had a potential relationship with jejunal VCR, plasma D-lactate and DAO, jejunal mucosa GSH-Px and CAT activity, and cecal short-chain fatty acid concentration. These data suggest that ED is effective in alleviating LPS-induced intestinal mucosal barrier injury by modulating gut microbiota in piglets.

Dietary Prebiotic Oligosaccharides and Arachidonate Alter the Fecal Microbiota and Mucosal Lipid Composition of Suckling Pigs

BackgroundEarly intestinal development is important to infant vitality, and optimal formula composition can promote gut health. ObjectivesThe objectives were to evaluate the effects of arachidonate (ARA) and/or prebiotic oligosaccharide (PRE) supplementation in formula on the development of the microbial ecosystem and colonic health parameters. MethodsNewborn piglets were fed 4 formulas containing ARA [0.5 compared with 2.5% of dietary fatty acids (FAs)] and PRE (0 compared with 8 g/L, containing a 1:1 mixture of galactooligosaccharides and polydextrose) in a 2 x 2 factorial design for 22 d. Fecal samples were collected weekly and analyzed for relative microbial abundance. Intestinal samples were collected on day 22 and analyzed for mucosal FAs, pH, and short-chain FAs (SCFAs). ResultsPRE supplementation significantly increased genera within Bacteroidetes and Firmicutes, including Anaerostipes, Mitsuokella, Prevotella, Clostridium IV, and Bulleidia, and resulted in progressive separation from controls as determined by Principal Coordinates Analysis. Concentrations of SCFA increased from 70.98 to 87.37 mM, with an accompanying reduction in colonic pH. ARA supplementation increased the ARA content of the colonic mucosa from 2.35–5.34% of total FAs. PRE supplementation also altered mucosal FA composition, resulting in increased linoleic acid (11.52–16.33% of total FAs) and ARA (2.35–5.16% of total FAs). ConclusionsPrebiotic supplementation during the first 22 d of life altered the gut microbiota of piglets and increased the abundance of specific bacterial genera. These changes correlated with increased SCFA, which may benefit intestinal development. Although dietary ARA did not alter the microbiota, it increased the ARA content of the colonic mucosa, which may support intestinal development and epithelial repair. Prebiotic supplementation also increased unsaturation of FAs in the colonic mucosa. Although the mechanism requires further investigation, it may be related to altered microbial ecology or biohydrogenation of FA.

In Vitro Effect on Piglet Gut Microbiota and In Vivo Assessment of Newly Isolated Bacteriophages against F18 Enterotoxigenic Escherichia coli (ETEC).

Enterotoxigenic Escherichia coli (ETEC) causing post-weaning diarrhea (PWD) in piglets have a detrimental impact on animal health and economy in pig production. ETEC strains can adhere to the host's small intestinal epithelial cells using fimbriae such as F4 and F18. Phage therapy could represent an interesting alternative to antimicrobial resistance against ETEC infections. In this study, four bacteriophages, named vB_EcoS_ULIM2, vB_EcoM_ULIM3, vB_EcoM_ULIM8 and vB_EcoM_ULIM9, were isolated against an O8:F18 E. coli strain (A-I-210) and selected based on their host range. These phages were characterized in vitro, showing a lytic activity over a pH (4-10) and temperature (25-45 °C) range. According to genomic analysis, these bacteriophages belong to the Caudoviricetes class. No gene related to lysogeny was identified. The in vivo Galleria mellonella larvae model suggested the therapeutic potential of one selected phage, vB_EcoS_ULIM2, with a statistically significant increase in survival compared to non-treated larvae. To assess the effect of this phage on the piglet gut microbiota, vB_EcoS_ULIM2 was inoculated in a static model simulating the piglet intestinal microbial ecosystem for 72 h. This study shows that this phage replicates efficiently both in vitro and in vivo in a Galleria mellonella model and reveals the safety of the phage-based treatment on the piglet microbiota.

Age Rather Than Supplementation with Oat β-Glucan Influences Development of the Intestinal Microbiota and SCFA Concentrations in Suckling Piglets.

The effects of early supplementation with oat β-glucan during the suckling period on piglet gut microbiota composition, concentrations of short-chain fatty acids, and gut physiological markers were assessed. Fifty piglets from five litters, balanced for sex and birth weight, were divided within litters into two treatment groups: β-glucan and control. Piglets in the β-glucan group received the supplement three times/week from day 7 of age until weaning. Rectal swab samples were collected from 10 piglets per treatment group (balanced across litters) from week 1 to week 4, and plasma samples were collected at 1, 3, and 4 weeks of age. Additional samples of intestinal tissues and jugular and portal vein plasma were collected from 10 animals at weaning (one per treatment group and litter). The concentrations of short-chain fatty acids in plasma and the microbiota composition in rectal swabs were mainly influenced by piglet age, rather than the supplement. There were significant differences in microbiota composition between litters and several correlations between concentrations of short-chain fatty acids in plasma and specific microbial taxa in rectal swabs. Overall, β-glucan supplementation did not have any clear impact on the gut environment in suckling piglets, whereas a clear age-related pattern emerged.

Maternal tributyrin supplementation in late pregnancy and lactation improves offspring immunity, gut microbiota, and diarrhea rate in a sow model.

Several studies have evaluated the effects of tributyrin on sow reproductive performance; however, none of these studies have investigated the effects of tributyrin on sow gut microbiota and its potential interactions with immune systems and milk composition. Therefore, we speculated that tributyrin, the combination of butyrate and mono-butyrin without odor, would reach the hindgut and affect the intestinal microbiota composition and play a better role in regulating sow reproductive performance, gut flora, and health. Thirty sows (Landrace × Yorkshire) were randomly divided into two groups: the control group (CON) and the tributyrin group (TB), which received basal diet supplemented with 0.05% tributyrin. The experimental period lasted for 35 days from late pregnancy to lactation. The results showed that TB supplementation significantly shortened the total parturition time and reduced the diarrhea rate in suckling piglets. On day 20 of lactation, the milk fat and protein levels increased by 9 and 4%, respectively. TB supplementation significantly improved the digestibility of dry material, gross energy, and crude fat in the sow diet, but had no significant effect on crude protein digestibility. Furthermore, TB supplementation increased the levels of IL-10, IL-6, and IgA in the blood of weaned piglets, but had no effect on maternal immunity. Analysis of the fecal microbial composition revealed that the addition of TB during late gestation and lactation increased the microbiota diversity in sows and piglets. At the phylum level, sows in the TB group had a slight increase in the relative abundance of Bacteroidota and Spirochaetota and a decrease in Firmicutes. At the order level, the relative abundance of Lactobacillales was increased in piglets and sows, and the TB group showed increased relative abundance of Enterobacterales and significantly decreased relative abundance of Oscillospirales in piglets. At family level, the relative abundance of Lactobacillaceae, Oscillospiraceae, and Christensenellaceae increased in sows, and the relative abundance of Enterobacteriaceae and Lactobacillaceae increased in piglets. At genus level, the relative abundance of Lactobacillus increased in sows and piglets, but the relative abundance of Subdoligranulum and Eubacterium_fissicatena_group decreased in piglets in the TB group. In conclusion, tributyrin supplementation shortened the farrowing duration and reduced the diarrhea rate of piglets by improving the inflammatory response and composition of gut microbiota in piglets and sows.

Impact of Intestinal Microbiota on Growth Performance of Suckling and Weaned Piglets.

Small-scale studies investigating the relationship between pigs' intestinal microbiota and growth performance have generated inconsistent results. We hypothesized that on farms under favorable environmental conditions (e.g., promoting sow nest-building behavior, high colostrum production, low incidence of diseases and minimal use of antimicrobials), the piglet gut microbiota may develop toward a population that promotes growth and reduces pathogenic bacteria. Using 16S rRNA gene amplicon sequencing, we sampled and profiled the fecal microbiota from 170 individual piglets throughout suckling and postweaning periods (in total 670 samples) to track gut microbiota development and its potential association with growth. During the suckling period, the dominant genera were Lactobacillus and Bacteroides, the latter being gradually replaced by Clostridium sensu scricto 1 as piglets aged. The gut microbiota during the nursery stage, not the suckling period, predicted the average daily growth (ADG) of piglets. The relative abundances of SCFA-producing genera, in particular Faecalibacterium, Megasphaera, Mitsuokella, and Subdoligranulum, significantly correlated with high ADG of weaned piglets. In addition, the succession of the gut microbiota in high-ADG piglets occurred faster and stabilized sooner upon weaning, whereas the gut microbiota of low-ADG piglets continued to mature after weaning. Overall, our findings suggest that weaning is the major driver of gut microbiota variation in piglets with different levels of overall growth performance. This calls for further research to verify if promotion of specific gut microbiota, identified here at weaning transition, is beneficial for piglet growth. IMPORTANCE The relationship between pigs' intestinal microbiota and growth performance is of great importance for improving piglets' health and reducing antimicrobial use. We found that gut microbiota variation is significantly associated with growth during weaning and the early nursery period. Importantly, transitions toward a mature gut microbiota enriched with fiber-degrading bacteria mostly complete upon weaning in piglets with better growth. Postponing the weaning age may therefore favor the development of fiber degrading gut bacteria, conferring the necessary capacity to digest and harvest solid postweaning feed. The bacterial taxa associated with piglet growth identified herein hold potential to improve piglet growth and health.

Bacillus amyloliquefaciens 40 regulates piglet performance, antioxidant capacity, immune status and gut microbiota.

Probiotics can improve animal growth performance and intestinal health. Bacillus species, Lactobacillus species, Bifidobacterium species, yeast etc. are the common types of probiotics. However, understanding the effects of probiotics on the immune status and gut microbiota of weaning piglets and how the probiotics exert their impact are still limited. This study aimed to investigate the effects of Bacillus amyloliquefaciens 40 (BA40) on the performance, immune status and gut microbiota of piglets. A total of 12 litters of newborn piglets were randomly divided into 3 groups. Piglets in control group were orally dosed with phosphate buffered saline; BA40 group and probiotics group were orally gavaged with resuspension BA40 and a probiotics product, respectively. The results showed that BA40 treatment significantly decreased (P<0.05) the diarrhea incidence (from d 5 to 40), diamine oxidase, D-lactate, interleukin (IL)-1β and interferon-γ concentrations compared with control group and probiotics group. Meanwhile BA40 dramatically increased the total antioxidant capacity, IL-10 and secretoryimmunoglobulin-A concentrations in contrast to control group. For the microbial composition, BA40 modulated the microbiota by improving the abundance of Bacteroides, Phascolarctobacterium (producing short-chain fatty acids) and Desulfovibrio and reducing the proliferation of pathogens (Streptococcus, Tyzzerella, Vellionella and paraeggerthella). Meanwhile, a metabolic function prediction explained that carbohydrate metabolism and amino acid metabolism enriched in BA40 group in contrast to control group and probiotics group. For correlation analysis, the results demonstrated that BA40-enriched Phascolarctobacterium and Desulfovibrio provide insights into strategies for elevating the health status and performance of weaned piglets. Altogether, BA40 exerted stronger ability in decreasing diarrhea incidence and improved antioxidant activity, gut barrier function and immune status of piglets than the other treatments. Our study provided the experimental and theoretical basis for the application of BA40 in pig production.

An insight into the commercial piglet’s microbial gut colonization: from birth towards weaning

BackgroundThe establishment of the gut microbiota can be influenced by several perinatal factors, including, most importantly, the maternal microbiota. Moreover, early-life environmental variation affects gut microbial colonization and the intestinal health of offspring throughout life. The present study aimed to explore the development of piglet gut microbiota from birth to weaning in the commercial practice and also to assess how different farm environments could condition this process. Although it is possible to find in the literature other studies with similar objectives this work probably represents one of the few studies that make a systematic evaluation of such differential factors under a real scenario. To achieve this objective, we performed two trials. In a first Trial, we selected 2 farms in which we performed an intensive sampling (5 samples /animal) to characterize the gut colonization pattern during the first days of life and to identify the time window with the greatest impact. Both farms differed in their health status and the use of antimicrobials in the piglets. In a second Trial, we selected 4 additional farms with variable rearing conditions and a distinctive use of antimicrobials in the sows with a simplified sampling pattern (2 samples/animal). Faecal samples were obtained with swabs and DNA was extracted by using the PSP® Spin Stool DNA Kit and sequencing of the 16S rRNA gene (V3-V4 region) performed by Illumina MiSeq Platform.ResultsThe present study contributes to a better understanding of microbiome development during the transition from birth to weaning in commercial conditions. Alpha diversity was strongly affected by age, with an increased richness of species through time. Beta diversity decreased after weaning, suggesting a convergent evolvement among individuals. We pinpointed the early intestinal colonizers belonging to Bacteroides, Escherichia-Shigella, Clostridium sensu stricto 1, and Fusobacterium genera. During lactation(d7-d21 of life), the higher relative abundances of Bacteroides and Lactobacillus genera were correlated with a milk-oriented microbiome. As the piglets aged and after weaning (d36 of life), increasing abundances of genera such as Prevotella, Butyricimonas, Christensenellaceae R-7 group, Dorea, Phascolarctobacterium, Rikenellaceae RC9 gut group, Subdoligranulum, and Ruminococcaceae UCG-002 were observed. These changes indicate the adaptation of the piglets to a cereal-based diet rich in oligosaccharides and starch. Our results also show that the farm can have a significant impact in such a process, evidencing the influence of different environments and rearing systems on the gut microbiota development of the young piglet. Differences between farms were more noticeable after weaning than during lactation with changes in alpha and beta biodiversity and specific taxa. The analysis of such differences suggests that piglets receiving intramuscular amoxicillin (days 2–5 of life) and being offered an acidifying rehydrating solution (Alpha farm in Trial 1) have a greater alpha diversity and more abundant Lactobacillus population. Moreover, the only farm that did not offer any rehydrating solution (Foxtrot farm in Trial 2) showed a lower alpha diversity (day 2 of life) and increased abundance of Enterobacteriaceae (both at 2 and 21 days). The use of in-feed antibiotics in the sows was also associated with structural changes in the piglets’ gut ecosystem although without changes in richness or diversity. Significant shifts could be registered in different microbial groups, particularly lower abundances of Fusobacterium in those piglets from medicated sows.ConclusionsIn conclusion, during the first weeks of life, the pig microbiota showed a relevant succession of microbial groups towards a more homogeneous and stable ecosystem better adapted to the solid dry feed. In this relevant early-age process, the rearing conditions, the farm environment, and particularly the antimicrobial use in piglets and mothers determine changes that could have a relevant impact on gut microbiota maturation. More research is needed to elucidate the relative impact of these farm-induced early life-long changes in the growing pig.

Effects of supplementing with Humulus scandens on the growth performance and gut microbiota in piglets.

This study was conducted to evaluate the effects of feeding Humulus scandens (Hu) on growth performance and gut microbiota in piglets. A total of 120 piglets were allocated to four dietary treatments (1) CON, basal diet; (2) T1, basal diet + 2.0% Hu; (3) T2, basal diet + 2.8% Hu and (4) T3, basal diet + 3.6% Hu. The results showed that dietary H. scandens supplementation increased the final body weight and average daily gain. Furthermore, H. scandens supplementation in T1 groups increased the content of total protein, globulin and IgG in serum and the apparent digestibility of crude protein. Gut microbiota analysis showed that H. scandens treatment in T1 groups increased the abundances of Lactobacillus, Ruminococcaceae, Enterococcus and Pseudomonas in cecum content. These findings suggested that dietary H. scandens supplementation improved the growth performance, immunological function and nutrient apparent digestibility as well as modulating the gut microbiota in piglets. This study contributed to developing new feed resources and might provide an alternative strategy for growth promotion in piglets.

Nourishing neonatal piglets with synthetic milk and Lactobacillus sp. at birth highly modifies the gut microbial communities at the post-weaning stage.

The importance of probiotics in pig production is widely recognized. However, the precise role of probiotics in regulating the gut microbiota of piglets has not been assessed extensively. Therefore, we intend to examine whether suckling pigs ingesting with synthetic milk (SM) and probiotics along with mother milk has a carryover effect on its growth and gut health at the post-weaning stage. A total of 40 [Duroc× (Yorkshire× Landrace)] neonates with an initial BW of 1.49 ± 0.28 kg were assigned to one of two treatments groups: control (CON) and treatment (TRT). Control group piglets were nourished with synthetic milk, while TRT group piglets were nourished SM with (1 × 109 CFU/g) Lactobacillus sp. probiotics. The treatment group piglets showed higher (p < 0.05) body weight and daily gain at week 3 than the CON group piglets. 16S metagenome sequencing showed average demultiplexed reads and denoised reads counts of 157,399 and 74,945, respectively. The total ASV taxonomy number classified with a confidence threshold > 70% (default) on sequence alignment with the SILVA v138 reference database was 4,474. During week 1, Escherichia-Shigella, Clostridium sensu stricto 1, and Bacteroides were confirmed as the major dominant bacterial genera in both the groups at the genus level. However, during week 2, the relative proportion of Escherichia-Shigella, Clostridium sensu stricto 1, and Proteobacteria was decreased, while that of Lactobacillus and Bacteroidota was increased in pigs receiving the probiotic supplement. During weeks 2 and 3, Firmicutes, Proteobacteria, and Bacteroidota phyla were dominant in both groups. During week 6, the relative proportion of Proteobacteria was slightly increased in both groups. Furthermore, Prevotella was confirmed as the major dominant bacterial genus in both groups during weeks 3 and 6. This study suggests that nourishing neonatal piglets with synthetic milk and Lactobacillus sp. probiotics from birth to 21 days would be beneficial to enhance the gut health of piglets and to overcome post-weaning mortality.

Effect of an Escherichia coli F4/F18 bivalent oral live vaccine on gut health and performance of healthy weaned pigs

Oral live vaccines stimulate host immunity, but they could also affect intestinal mucosa development and gut microbiota of piglets during the postweaning. The aim of this study was to determine the effect of an oral vaccine against Escherichia coli F4 and F18 (Coliprotec F4/F18®), on gut functionality and integrity, growth performance and health status of postweaning piglets. A total of 96 weaned piglets (23.30 ± 1.85 days of age; 7334 ± 1039 g BW) were divided into two groups (16 replicates/group; three piglets/replicate) as follows: (1) Control (CO), fed a standard diet (prestarter up to 14 days, then starter feed); (2) Treated (TRT): as CO but vaccinated with Coliprotec F4/F18® at weaning (day 0). Piglets were weighed at day 0 and weekly until day 35. Individual faecal score was recorded daily. Piglets were sacrificed at days 10 (1/3 of total) and 35 (2/3). Samples of jejunum mucosa and of cecum content were collected for morphometric, immunohistochemistry analysis and for microbiota profile analysis, respectively. Data were fitted using a linear model including treatment, class of starting BW as fixed factors and litter as random factor. From days 0 to 7, piglets from the TRT group tended to have a higher average daily gain (+22.6%, P = 0.08) and average daily feed intake compared to the CO group (+13.2%, P = 0.022). Gain to feed ratio was lower in the TRT group from days 14 to 35 (−6.6%, P = 0.011). From days 7 to 14, the TRT group had a higher diarrhoea index (−199%, P < 0.001). Crypt depth was higher in the CO group (+10.9%, P = 0.04) at day 10, but not at day 35. Jejunal expression of Claudin-4 (probability of having a score = 3) was higher in the TRT group at day 10 (CO = 1.50% vs TRT = 2.69%, P < 0.0001) and day 35 (CO = 1.29% vs TRT = 1.92%, P = 0.012). Oral vaccine affected beta diversity at day 10 (P = 0.040; R2 = 0.05) and increased the abundance of specific taxa and genera in the cecum at day 10, including Prevotella (lg2FC = 23.2, FDR < 0.001). The results showed how an Escherichia coli-based vaccine supplied to weaned pigs can promote gut health by controlling symptoms of the postweaning perturbation in the first 2 weeks postweaning. In addition, the vaccine strains showed a probiotic-like effect by modulating gut microbiota favouring the establishment of beneficial bacteria, and by promoting gut barrier integrity.

Gut microbiota and growth performance of offspring are influenced by wet nurse in pigs using cross-fostering trial.

Nursing mom can regulate the gut microbiome succession in offspring. However, it remains unclear whether these effects are long-term and what effect it has on the growth performance of piglets. This study aimed to develop a cross-fostering model of piglets and investigate the effect of maternal environment on gut microbiota, even the growth performance of the offspring, and if this effect could be maintained in the long term. Four groups of piglets were generated as follows (n= 12): Duroc piglets nursed by their birth mom (Dd) or Yorkshire sows (Yd), Yorkshire piglets nursed by Duroc sows (Dy) or their birth mom (Yy). The study found that cross-fostering improved the growth performance of piglets for a long time. The gut microbiota of piglets was mainly determined by the breeds of nursing moms before weaning, and it was more and more influenced by their breeds after weaning, but the influence of birth mom breeds still existed. The linear discriminant analysis (LDA) effect size (LEfSe) analysis and Spearman correlation analysis showed that Sutterella, Butyricimonas and Alistipes, which were affected by nursing mom before weaning, had a strong positive correlation with the growth performance of piglets before weaning. Candidatus_Soleaferrea and Treponema, which were affected by both nursing mom and piglet breed after weaning, were significantly negatively correlated with the growth performance of piglets long after weaning. Our results revealed that both the breeds of piglets and their birth moms influence the gut microbiota of piglets for a long time, even after weaning. Additionally, this effect might be related to the growth performance of piglets. © 2022 Society of Chemical Industry.